Cell-free synthesis of actin-like cytoskeleton filaments

MreB, an actin-like cytoskeleton in E. coli encoded by MreB gene, is essential for the maintenance of cellular shape of non-spherical bacteria. Our project is aimed at cell-free synthesis of actin homolog MreB, which is further characterized through both microfluidics technology and nanobiotechnology. As in vivo, MreB is rigidly linked to the cell wall. Here, MreB was cell-free expressed in vitro. Protein localization on the inner oil film could be observed in W/O/W microdroplets produced with microfluidic, which mimic cell membrane to provide a non-polar environment. Furthermore, SWNTs (single-walled carbon nanotubes) were involved as scaffolds for auxiliary aggregation of actin homolog MreB. The combination of MreB with SWNT was observed under AFM (atomic force microscope). It seems that aggregated MreB winded on the carbon nanotubes. In conclusion, we synthesized the actin-like cytoskeleton filaments in vitro, and provided a sound microenvironment for MreB synthesis, which might contribute to recognizing, exploring and even creating lives.